Imperial College London
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Emeritus Professor David Lane

Faculty of Medicine, Department of Immunology and Inflammation

Emeritus Professor of Haematology
 
 
 
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Contact

 

+44 (0)20 3313 2295d.lane

 
 
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Location

 

Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Santamaria:2019:10.1038/s41598-019-47494-w,
author = {Santamaria, S and Yamamoto, Y and Teraz-Orosz, A and Koch, C and Apte, SS and de, Groot R and Lane, DA and Ahnstroem, J},
doi = {10.1038/s41598-019-47494-w},
journal = {Scientific Reports},
title = {Exosites in hypervariable loops of ADAMTS dpacer domains control substrate recognition and proteolysis},
url = {http://dx.doi.org/10.1038/s41598-019-47494-w},
volume = {9},
year = {2019}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - ADAMTS (A Disintegrin-like and Metalloproteinase domain with Thrombospondin type 1 Motif)-1, -4 and -5 share the abilities to cleave large aggregating proteoglycans including versican and aggrecan. These activities are highly relevant to cardiovascular disease and osteoarthritis and during development. Here, using purified recombinant ADAMTS-1, -4 and -5, we quantify, compare, and define the molecular basis of their versicanase activity. A novel sandwich-ELISA detecting the major versican cleavage fragment was used to determine, for the first time, kinetic constants for versican proteolysis. ADAMTS-5 (kcat/Km 35 × 105 M−1 s−1) is a more potent (~18-fold) versicanase than ADAMTS-4 (kcat/Km 1.86 × 105 M−1 sec−1), whereas ADAMTS-1 versicanase activity is comparatively low. Deletion of the spacer domain reduced versicanase activity of ADAMTS-5 19-fold and that of ADAMTS-4 167-fold. Co-deletion of the ADAMTS-5 cysteine-rich domain further reduced versicanase activity to a total 153-fold reduction. Substitution of two hypervariable loops in the spacer domain of ADAMTS-5 (residues 739–744 and 837–844) and ADAMTS-4 (residues 717–724 and 788–795) with those of ADAMTS-13, which does not cleave proteoglycans, caused spacer-dependent reductions in versicanase activities. Our results demonstrate that these loops contain exosites critical for interaction with and processing of versican. The hypervariable loops of ADAMTS-5 are shown to be important also for its aggrecanase activity. Together with previous work on ADAMTS-13 our results suggest that the spacer domain hypervariable loops may exercise significant control of ADAMTS proteolytic activity as a general principle. Identification of specific exosites also provides targets for selective inhibitors.
AU  - Santamaria,S
AU  - Yamamoto,Y
AU  - Teraz-Orosz,A
AU  - Koch,C
AU  - Apte,SS
AU  - de,Groot R
AU  - Lane,DA
AU  - Ahnstroem,J
DO  - 10.1038/s41598-019-47494-w
PY  - 2019///
SN  - 2045-2322
TI  - Exosites in hypervariable loops of ADAMTS dpacer domains control substrate recognition and proteolysis
T2  - Scientific Reports
UR  - http://dx.doi.org/10.1038/s41598-019-47494-w
UR  - http://hdl.handle.net/10044/1/72307
VL  - 9
ER  -
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